The Massachusetts and Yale ADRC Collaborative Proteomic Biofluid Biomarker Discovery Program

目前还没有阿尔茨海默病(AD)的实用生物标志物，可以常规地 分期、评估预后和监测治疗反应。AD是一种复杂的疾病 重叠的病理生理学导致最终的突触丢失和进行性痴呆。 脑脊液中AD核心生物标志物淀粉样蛋白-b和tau(总磷)的测定 液体(脑脊液)，提供了有或没有实质性AD病理的强烈指示， 但还不能很好地反映疾病的分期或进展。为了更早地诊断出这种疾病， 了解并发病理生理学的时间进程，并最大限度地发挥 随着AD的分期、追踪和治疗，更多有用的AD生物标志物的特征是至关重要的。 耶鲁大学和马萨诸塞州ADRC之间提出了一项合作，将利用 这两个中心发现通路驱动的AD新生物标记物的个体优势。 使用大量具有临床特征的生物流体样本，这些样本来自 马萨诸塞州ADRC，耶鲁ADRC将利用他们的专业知识在定量目标LC- MS/MS技术可以评估脑脊液中的100多种分析物，并扩展到最多 对血液来说是有用的。LC-MS/MS具有固有的专属性，可同时检测 许多标记物来自一个小的生物液体体积。使用数据独立采集(DIA)LC- MS/MS技术允许重新分析原始数据，以便对新的 生物标志物，提供了一个可重复但灵活的发现管道。 在目标1中，脑脊液中的100多种蛋白质来自参与AD的通路 病理生理学将接受全面的技术资格审查。使用来自 作为临床试验的安慰剂，多肽将进行线性、批间、批内测试 和短期的生物时间稳定性。然后，所有高效的多肽都将在 60名认知正常成人(CU-A)和60名阿尔茨海默病(AD)痴呆患者的高对比样本 案子。他们将被评估他们区分AD和CU-A的能力，以及他们的 与突触功能标志物的相关性。在目标2中，技术合格的多肽将是 在3个量身定制的样本集合中进行量化，以评估他们在AD分期、预后方面的表现 和鉴别诊断。在目标3中，来自目标1和目标2的信息量最大的分析物将是 通过LC-MS/MS，或通过灵敏或超灵敏的ELISA法，在血液中进行定量。 将比较不同生物体液中的多肽丰度，以及这些标记物的诊断效用 在血液检测中。

There are currently no practical biomarkers for Alzheimer’s Disease (AD) that can routinely stage, assess prognosis, and monitor treatment response. AD is a complex disease of overlapping pathophysiologies that lead to eventual synapse loss and progressive dementia. The core AD biomarkers, amyloid-b and tau (total and phospho-) measured in cerebrospinal fluid (CSF), provide a strong indication of the presence or absence of substantial AD pathology, but as yet poorly reflect disease stage or progression. In order to diagnose the disease earlier, learn about the time-course of concurrent pathophysiologies, and maximize the potential for staging, tracking and treatment, it is critical that more useful AD biomarkers are characterized. A collaboration is proposed between the Yale and Massachusetts ADRCs that will leverage the individual strengths of these two centers to discover pathway driven novel biomarkers of AD. Using the large numbers of clinically characterized biofluid samples available from the Massachusetts ADRC, the Yale ADRC will employ their expertise in quantitative targeted LC- MS/MS technology to assess over one hundred analytes in CSF, with an extension of the most informative to blood. LC-MS/MS is inherently specific, and enables simultaneous testing of many markers from a small biofluid volume. Use of Data-Independent-Acquisition (DIA) LC- MS/MS techniques allows for primary data to be reanalyzed for retrospective analysis of new biomarkers, providing a reproducible yet flexible discovery pipeline. In Aim 1, a panel of over 100 proteins in CSF from pathways involved in AD pathophysiology will be subjected to comprehensive technical qualification. Using samples from the placebo arm of a clinical trial, peptides will be tested for linearity, inter-assay, intra-assay and short term biotemporal stability. All high performing peptides will then be quantified in a high contrast sample of 60 Cognitively unimpaired-adults (CU-A) and 60 dementia due to AD cases. They will be assessed for their ability to distinguish AD from CU-A, and for their correlation with markers of synaptic function. In Aim 2, technically qualified peptides will be quantified in 3 tailored sample collections to assess their performance in AD staging, prognosis and differential diagnosis. In Aim 3, the most informative analytes from Aims 1 & 2 will be quantified in blood, either by LC-MS/MS, or by sensitive or ultra-sensitive ELISA approaches. Peptide abundance will be compared across biofluids, and the diagnostic utility of these markers in blood assessed.